Development of tools to explore the fundamentals of axonemal dynein heavy chain biology and improve the efficiency of genome editing

Motile cilia are microtubule based projections that assist in the movement of fluid over the surface of cells, such as in the respiratory epithelium, or of cells through a fluid, such as in sperm. Ciliary movement is driven by axonemal dyneins (ADs), large molecular complexes which contain long heavy chain ATPase motor subunits. The stability of ADs has been shown to be dependent on multiple cytoplasmically localised proteins, which are involved in their assembly and trafficking to the cilia. The heavy chain subunits have been suggested to be particularly reliant on specialised chaperoning pathways in order to fold into the correct tertiary structure. Hereditary defects in genes encoding the proteins of the ADs or proteins involved in their assembly result in an incurable human disease, primary ciliary dyskinesia (PCD). PCD results in neonatal respiratory distress with lifelong respiratory complications and is also highly heterogenous with mutations in 40 genes associated with it so far. Despite the identification of many putative assembly factors, where and how they interact with AD proteins remains unknown. In order to investigate AD complexes, from the translation of their subunits to their degradation, in greater spatial and temporal detail a heavy chain outer dynein arm subunit (ODA), Dnah5, was tagged with the adaptable SNAP tag in mice. Dnah5 is the largest AD heavy chain and the most commonly mutated gene in PCD. When developing novel therapeutics the SNAP-Dnah5 mouse could be used as a reporter for functional rescue in PCD mouse models which exhibit loss of these complexes from the cilia. The effectiveness of the therapy could then be graded on the restoration of SNAP-DNAH5 fluorescence in the motile cilia. As a secondary aim this project also sought to improve the efficiency of CRISPR/Cas9 induced gene correction, via a novel linkage method, to develop a genome editing therapy for PCD, which could be tested using SNAP-Dnah5 mice. Using the SNAP-Dnah5 mouse tracheal epithelial cells I have directly imaged DNAH5’s docking onto the motile axoneme from the distal end and have demonstrated that there is a very low level of ODA turnover in mature cilia. I have also shown that the Dnah5 transcript localises to large apical clusters in ciliated tracheal epithelial cells and via preliminary pulldown experiments that SNAPDNAH5 might interact with RNA regulatory proteins in maturing motile ciliated cells suggestive of translational regulation. This project demonstrates the utility of this mouse model for future studies

Inequalities and inclusion in exercise referral schemes: a mixed-method multi-scheme analysis

Physical activity prescription, commonly through exercise referral schemes, is an established disease prevention and management pathway. There is considerable heterogeneity in terms of uptake, adherence, and outcomes, but because within-scheme analyses dominate previous research, there is limited contextual understanding of this variance. Both the impact of schemes on health inequalities and best practices for inclusion of at-risk groups are unclear. To address this, we modelled secondary data from the multi-scheme National Referral Database, comprising 23,782 individuals across 14 referral schemes, using a multilevel Bayesian inference approach. Scheme-level local demographics identified over-sampling in uptake; on the basis of uptake and completion data, more inclusive schemes (n = 4) were identified. Scheme coordinators were interviewed, and data were analyzed using a grounded theory approach. Inequalities presented in a nuanced way. Schemes showed promise for engaging populations at greater risk of poor health (e.g., those from more deprived areas or of an ethnic minority background). However, the completion odds were lower for those with a range of complex circumstances (e.g., a mental health-related referral). We identified creative best practices for widening access (e.g., partnership building), maintaining engagement (e.g., workforce diversity), and tailoring support, but recommend changes to wider operational contexts to ensure such approaches are viable

We are failing to improve the evidence base for “Exercise Referral”: How a Physical Activity Referral Scheme Taxonomy can help

Abstract not available

A modified Delphi study to gain consensus for a taxonomy to report and classify physical activity referral schemes (PARS)

Background: Physical Activity Referral Schemes (PARS), including exercise referral schemes, are a popular approach to health improvement, but understanding of effectiveness is limited by considerable heterogeneity in reporting and evaluation. We aimed to gain consensus for a PARS taxonomy as a comprehensive method for reporting and recording of such schemes. Methods: We invited 62 experts from PARS policy, research and practice to complete a modified Delphi study. In round one, participants rated the need for a PARS taxonomy, the suitability of three proposed classification levels and commented on proposed elements. In round two, participants rated proposed taxonomy elements on an 11-point Likert scale. Elements scoring a median of ≥7, indicating high agreement, were included in the final taxonomy. Results: Of those invited, 47 (75.8%) participated in round one, with high retention in round two (n = 43; 91.5%). 42 were UK-based, meaning the resultant taxonomy has been scrutinised for fit to the UK context only. The study gained consensus for a three-level taxonomy: Level 1: PARS classification (primary classification, provider, setting, conditions accepted [have or at risk of], activity type and funding). Level 2: scheme characteristics (staff structure, staff qualifications, behaviour change theories, behaviour change techniques, referral source, referrers, referral process, scheme duration, session frequency, session length, session times, session type, exit routes, action in case of non-attendance, baseline assessment, exit assessment, feedback to referrer and exclusion criteria) and Level 3: participant measures (demographics, monitoring and evaluation, and measures of change). Conclusion: Using a modified Delphi method, this study developed UK-based consensus on a PARS classification taxonomy. We encourage PARS practitioners and public health colleagues, especially those working with similar service models internationally, to test, refine and use this taxonomy to inform policy and practice

A scoping review of methods and measures used to capture children's play during school breaktimes

Play is linked to healthy child development and is recognised in the UN Convention on the Rights of the Child. School breaktimes provide regular opportunities for children to play, and as such, they have been the context of a large and interdisciplinary body of research on play. Play research has diverse aims and cuts across many academic disciplines, resulting in a wide range of methods and measurement tools being used in research to capture children’s play. In this scoping review, 105 studies of play during school breaktimes were identified and we describe, synthesise and compare methods used to assess play during school breaktimes, bringing together methodologies from different fields for the first time. Specifically, we captured: the aspects of play that have been measured and described; established tools and coding schemes that have been used; what the measures of play have been used for; and what the quality of reporting of play measures has been. In this way, we anticipate that the review will facilitate future play research and support, where appropriate, more consistent use and transparent reporting of methods and measures

Revision of EU Ecolabel Criteria for Electronic Displays (previously Televisions). Final Technical Report: Final criteria

This technical report is aimed at providing a sound base to the revision process of the EU Ecolabel criteria for Televisions. Currently, two separate sets of EU Ecolabel criteria exist for televisions (Commission Decision 2009/300/EC) and for external computer displays as part of the criteria set for personal computers (Commission Decision 2011/337/EU). The revised Ecolabel criteria document is proposed to cover both product groups; thus common criteria for both televisions and external computer displays will be developed, differentiating between technical product characteristics where necessary. The product groups has been renamed as Electronic Displays. To achieve those objectives, a Preliminary Report was produced together with the first version of this technical report as a working document. Hence, this document was developed to undergo the stakeholder consultation, which is crucial to come up with criteria adapted to the market reality while being able to select the best environmental performance products available on the market. After an extensive stakeholder's consultation process,it has been produced the this final version of the Technical Report supporting the final criteria for electronic displays.JRC.B.5-Circular Economy and Industrial Leadershi

Parent perceived barriers and facilitators of children's adventurous play in Britain: a framework analysis

This is the final version. Available on open access from BMC via the DOI in this recordAvailability of data and materials: The data obtained and analysed in the current study are available via the following link: https://doi.org/10.5255/UKDA-SN-8793-1.BACKGROUND: From a public health perspective there is growing interest in children's play, including play involving risk and adventure, in relation to children's physical and mental health. Regarding mental health, it is theorised that adventurous play, where children experience thrilling, exciting emotions, offers important learning opportunities that prepare children for dealing with uncertainty and help prevent anxiety. Despite these benefits, adventurous play has decreased substantially within a generation. Parents have a key role in facilitating or limiting children's opportunities for adventurous play, but research identifying the barriers and facilitators parents perceive in relation to adventurous play is scarce. The present study therefore examined the barriers to and facilitators of adventurous play as perceived by parents of school-aged children in Britain. METHODS: This study analysed data from a subsample of parents in Britain (n = 377) who participated in the nationally representative British Children's Play Survey. Parents responded to two open-ended questions pertaining to the barriers to and facilitators of children's adventurous play. Responses were analysed using a qualitative Framework Analysis, an approach suitable for managing large datasets with specific research questions. RESULTS: Four framework categories were identified: Social Environment; Physical Environment; Risk of Injury; Child Factors. Social Environment included barriers and facilitators related to parents, family and peers, as well as community and society. Dominant themes within the Social Environment related to perceptions about the certainty of child safety, such as supervision and the safety of society. Beliefs about the benefits of adventurous play for development and well-being were also important in the Social Environment. Physical Environment factors focused on safety and practical issues. Risk of Injury captured concerns about children being injured during play. Child Factors included child attributes, such as play preference, developmental ability and trait-like characteristics. CONCLUSIONS: Improved understanding of what influences parent perceptions of adventurous play can inform public health interventions designed to improve children's opportunities for and engagement in adventurous play, with a view to promote children's physical and mental health.Economic and Social Research Council (ESRC)UKR

Physical activity for general health benefits in disabled children and disabled young people: rapid evidence review<strong> </strong>

Executive summary: Disabled children and disabled young people are at increased risk of being typically inactive, particularly as they become older. These issues have been exacerbated by the COVID-19 pandemic. This public health-focused report provides a review of the scientific evidence of the health benefits from physical activity, specifically for disabled children and disabled young people in non-clinical settings (aged 2 to 17 years). The research responds to a gap identified in the review of evidence for the 2019 UK Chief Medical Officers’ (CMOs’) physical activity guidelines. This important adjunct report enhances the comprehensiveness of the UK’s physical activity guidelines provision. It also provides future research recommendations. Furthermore, the report will also inform the first evidence-based infographic co-produced with disabled children and disabled young people, their parents and carers, health and social care professionals, and key disability and sport organisations to communicate meaningful messages about these physical activity recommendations, especially in relation to the frequency and duration of activity.There is evidence that shows a likely relationship between engaging in physical activity and positive health outcomes for disabled children and disabled young people. This report also provides suggestions about the amount (that is, frequency, duration and intensity) of physical activity that is likely to be important for disabled children and disabled young people to undertake to benefit their health. Some physical activity is better than nothing, as small amounts can bring health benefits. For likely substantial health gains, it is important for disabled children and disabled young people to do 120 to 180 minutes of mostly aerobic physical activity per week, at a moderate-to-vigorous intensity. The evidence suggests that this may be achieved in different ways (for example, 20 minutes per day or 40 minutes 3 times per week). It is also important for disabled children and disabled young people to do challenging strength and balance-focused activities on average 3 times per week. No evidence was found to show that physical activity is unsafe for disabled children and disabled young people when it is performed at an appropriate level for their current levels of physical development, fitness, physical and mental functioning (accounting for disability classification and severity), health and physical activity.This report provides evidence that aligns in part with the 2019 UK Chief Medical Officers’ physical activity guidelines for non-disabled children and disabled young people, as well as the 2020 guidelines published by the World Health Organization (WHO) for disabled children and disabled young people. However, there are also important differences in terms of recommended frequency and time. These are made based on the available evidence to provide recommendations specific to disabled children and disabled young people. The report also aids the communication and implementation of the guidelines by providing an evidence-base for disability groups, health and social care professionals, and sport and physical activity organisations to encourage physical activity to disabled children and disabled young people.The guidelines are the first to have included a review of evidence solely focused on disabled children and disabled young people’s physical activity, and thus represent the most comprehensive guidance globally

Hyperpolarized 13 C and 31 P MRS detects differences in cardiac energetics, metabolism, and function in obesity, and responses following treatment

Obesity is associated with important changes in cardiac energetics and function, and an increased risk of adverse cardiovascular outcomes. Multi‐nuclear MRS and MRI techniques have the potential to provide a comprehensive non‐invasive assessment of cardiac metabolic perturbation in obesity. A rat model of obesity was created by high‐fat diet feeding. This model was characterized using in vivo hyperpolarized [1‐13C]pyruvate and [2‐13C]pyruvate MRS, echocardiography and perfused heart 31P MRS. Two groups of obese rats were subsequently treated with either caloric restriction or the glucagon‐like peptide‐1 analogue/agonist liraglutide, prior to reassessment. The model recapitulated cardiovascular consequences of human obesity, including mild left ventricular hypertrophy, and diastolic, but not systolic, dysfunction. Hyperpolarized 13C and 31P MRS demonstrated that obesity was associated with reduced myocardial pyruvate dehydrogenase flux, altered cardiac tricarboxylic acid (TCA) cycle metabolism, and impaired myocardial energetic status (lower phosphocreatine to adenosine triphosphate ratio and impaired cardiac ΔG~ATP). Both caloric restriction and liraglutide treatment were associated with normalization of metabolic changes, alongside improvement in cardiac diastolic function. In this model of obesity, hyperpolarized 13C and 31P MRS demonstrated abnormalities in cardiac metabolism at multiple levels, including myocardial substrate selection, TCA cycle, and high‐energy phosphorus metabolism. Metabolic changes were linked with impairment of diastolic function and were reversed in concert following either caloric restriction or liraglutide treatment. With hyperpolarized 13C and 31P techniques now available for human use, the findings support a role for multi‐nuclear MRS in the development of new therapies for obesity

Adventurous play for a healthy childhood: facilitators and barriers identified by parents in Britain

Rationale Adventurous play, where children take age-appropriate risks involving uncertainty, fear, and thrill, is positively associated with children's physical health, mental health, and development. There is growing concern that children's access to and engagement with adventurous play opportunities are declining in Westernised countries, which may have negative implications for children's health. Objective The current study aimed to ascertain the facilitators of and barriers to children's adventurous play most identified by parents in Britain and to determine whether these differ across socio-demographic and geographic groups. Methods This study analysed the responses of a nationally representative sample of 1919 parents who took part in the British Children's Play Survey. Two open-ended questions asked parents to identify what they perceive to be the facilitators of and barriers to their child's adventurous play. A quantitative coding scheme, developed using the qualitative framework identified by Oliver et al. (2022), was applied to parents' responses. Results A diversity in the most identified facilitators and barriers was found, including concerns about the risk of injury from adventurous play and the safety of society, positive attitudes about the benefits of adventurous play, as well as factors related to child attributes. In general, these were consistently identified across different socio-demographic and geographic groups, although some differences were found in barriers. Conclusions The findings of this research support the identification of key targets for those working with parents to improve children's adventurous play opportunities and ultimately their physical and mental health. Future research should seek to design and tailor interventions by asking parents about the support they would value